Methods of using IL-1 antagonists to treat autoinflammatory disease

ABSTRACT

Methods of treating, inhibiting, or ameliorating an autoinflammatory disorder, disease, or condition in a subject in need thereof, comprising administering to a subject in need a therapeutic amount of an interleukin 1 (IL-1) antagonist, wherein the autoinflammatory disorder, disease, or condition is treated, inhibited, or ameliorated. The IL-1 antagonist is a molecule capable of binding and inhibiting IL-1. The therapeutic methods are useful for treating a human adult or child suffering from Neonatal Onset Multisystem Inflammatory Disorder (NOMID/CINCA), Muckle-Wells Syndrome (MWS), Familial Cold Autoinflammatory Syndrome (FCAS), familial mediterranean fever (FMF), tumor necrosis factor receptor-associated periodic fever syndrome (TRAPS), or systemic onset juvenile idiopathic arthritis (Still&#39;s Disease).

RELATED APPLICATIONS

This application claims the benefit under 35 USC 119(e) to U.S. Ser. No.60/577,023 filed 4 Jun. 2004, which application is incorporated byreference in its entirety.

BACKGROUND

1. Field of the Invention

The invention relates to methods of using interleukin-1 (IL-1)antagonists to treat autoinflammatory diseases, such as, for example,including familial mediterranean fever (FMF), NOMID/CINCA, Muckle-WellsSyndrome, FCAS, and tumour necrosis factor receptor-associated periodicfever syndrome (TRAPS).

2. Description of Related Art

One important group of autoinflammatory disorders encompasses autosomaldominant conditions associated with mutations in CIAS-1, a gene thatencodes a pyrin-related protein called “cryopyrin” (Feldmann et al.(2002) Am. J. Hum. Genet. 71:198-203; Hoffman et al. (2001) Nat. Genet.29:301-305). These disorders include Neonatal Onset MultisystemInflammatory Disorder (NOMID/CINCA), Muckle-Wells Syndrome (MWS), andFamilial Cold Autoinflammatory Syndrome (FCAS). These disorders presenta spectrum of clinical manifestations ranging from FCAS being themildest to the seriously disabling disease of NOMID/CINCA. Anurticaria-like skin rash is common to the entire spectrum of thesediseases. In patients with FCAS, this rash is inducible by cold exposurewhile most patients with MWS or NOMID present with daily rashes that areconsistently provoked by a number of different stimuli. Conjunctivitisis present in all forms of disease expression, however, hearing loss,aseptic meningitis and arthritis are mainly seen in patients with MWSand NOMID/CINCA. The disfiguring and disabling body overgrowth at theepiphyses and patellae is only seen in patients with NOMID/CINCA.

FMF is a recessively inherited condition characterized by episodes offever and serositis or synovitis; some subjects also develop systemicamyloidosis (Balow et al. (1997) Genomics 44:280-291). The FMF geneencodes a novel protein called pyrin that is the prototype of a familyof molecules involved in the regulation of apoptosis (cell-death) andinflammation. The precise biochemical mechanism by which these proteinsfunction, and by which mutations cause disease, is still unknown.

Still's Disease (systemic onset juvenile idiopathic arthritis), ismanifest by spiking fevers, evanescent salmon color rash, arthritis,arthralgia, and hepatosplenomegaly (Masson et al. (1995) Rev. Rhum.Engl. Ed. 62:748-757; Spiegel et al. (2000) Arthritis Rheum.43:2402-2409). There are as yet no definitive genetic associations withStill's Disease and the pathogenesis is poorly understood.Interestingly, many of the signs and symptoms of Still's disease aresimilar to those with autoinflammatory disease. Still's Diseasetypically first occurs during childhood, but can also have its onset inadulthood.

Similarly, Kawasaki disease is a disease affecting children that isaccompanied by fevers, swelling and arthritic joints, and rash, as wellas vascular inflammation that can cause permanent coronary damage inapproximately 15-25% of affected children. Two other similar diseasesare Blau's syndrome and Early Onset Sarcoidosis (EOS), both of which arecaused by a gain of function mutations in NOD2, a protein similar toPyrin, and cause rash, granulomatosis, arthritis and uveitis. Otherdiseases that have also been considered autoinflammatory include,Hidradenitis suppurativa, Behcet's, hyperimmunoglobulinemia D withperiodic fever syndrome (HIDS), tumour necrosis factorreceptor-associated periodic fever syndrome (TRAPS), and Pyogenicsterile arthritis, pyoderma gangrenosum and acne (PAPA syndrome).

The pathogenesis of autoinflammatory disease is not completelyunderstood. There is a growing body of evidence that interleukin-1(IL-1) plays a role in a number of these conditions and that targetingof this cytokine can provide important benefits (Hoffman et al. (2004)Arthritis. Rheum. 50:345-349). There is clearly a need to developimproved therapeutic treatment of these autoinflammatory diseases.

BRIEF SUMMARY OF THE INVENTION

In a first aspect, the invention features a method of treating,inhibiting, or ameliorating an autoinflammatory disorder, comprisingadministering to a subject in need an interleukin 1 (IL-1) antagonist.An IL-1 antagonist is a compound capable of blocking or inhibiting thebiological action of IL-1, including IL-1-binding fusion proteins. In apreferred embodiment, the IL-1 antagonist is an IL-1-specific fusionprotein comprising two IL-1 receptor components and a multimerizingcomponent, for example, an IL-1 fusion protein trap antagonist (an “IL-1trap”) described in U.S. patent publication No. 2003/0143697, published31 Jul. 2003, herein specifically incorporated by reference in itsentirety. In a specific embodiment, the IL-1 antagonist is the fusionprotein shown in SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26.A preferred fusion protein is shown in SEQ ID NO:10. The inventionencompasses the use of an IL-1-binding fusion protein substantiallyidentical to the protein of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20,22, 24, 26, that is, a protein having at least 95% identity, at least97% identity, at least 98% identity to the protein of SEQ ID NO: 4, 6,8, 10,12, 14, 16, 18, 20, 22, 24, 26 and capable of binding andinhibiting IL-1. Further, in specific embodiments, the IL-1 antagonistis a fusion protein comprising one or more immunoglobulin-derivedcomponents in place of one or more receptor components. In specificembodiments, the IL-1 antagonist comprises one or moreimmunoglobulin-derived components specific for IL-1 and/or an IL-1receptor.

The subject being treated is most preferably a human diagnosed assuffering from an autoinflammatory disorder. More specifically, thesubject is a human adult or child diagnosed with an autoinflammatorydisorder associated with mutations in CIAS-1, such as Neonatal OnsetMultisystem Inflammatory Disorder (NOMID/CINCA), Muckle-Wells Syndrome(MWS), Familial Cold Autoinflammatory Syndrome (FCAS); familialMediterranean fever (FMF); systemic onset juvenile idiopathic arthritis(Still's Disease), tumour necrosis factor receptor-associated periodicfever syndrome (TRAPS), or Kawasaki Disease.

The method of the invention includes administration of the IL-1antagonist by any means known to the art, for example, subcutaneous,intramuscular, intranasal, intravenous, transdermal administration ororal routes of administration. Preferably, administration issubcutaneous or intravenous.

In a second aspect, the invention features a method of treating,inhibiting, or ameliorating a disease or condition selected from thegroup consisting of NOMID/CINCA, MWS, FCAS, FMP, Still's Disease, TRAPS,and Kawasaki Disease, the method comprising administering to a subjectin need an interleukin 1 (IL-1) antagonist. In a preferred embodiment,the IL-1 antagonist is a fusion protein capable of trapping IL-1. In aspecific embodiment, the IL-1 antagonist is the fusion protein shown inSEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, or asubstantially identical protein capable of binding and inhibiting IL-1.A preferred IL-1 antagonist is shown in SEQ ID NO:10. Preferably, thesubject treated is a child or adult human diagnosed with the disease orcondition.

In a third aspect, the invention features a method of treating,inhibiting, or ameliorating Neonatal Onset Multisystem InflammatoryDisorder (NOMID/CINCA), comprising administering to a subject in need aninterleukin 1 (IL-1) antagonist. In a preferred embodiment, the IL-1antagonist is a fusion protein capable of trapping IL-1. In a specificembodiment, the IL-1 antagonist is the fusion protein shown in SEQ IDNO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, or a substantiallyidentical protein capable of binding and inhibiting IL-1. A preferredIL-1 antagonist is shown in SEQ ID NO:10.

In a fourth aspect, the invention features a method of treating,inhibiting, or ameliorating Muckle-Wells Syndrome (MWS), the methodcomprising administering to a subject in need an interleukin 1 (IL-1)antagonist. In a preferred embodiment, the IL-1 antagonist is a fusionprotein capable of trapping IL-1. In a specific embodiment, the IL-1antagonist is the fusion protein shown in SEQ ID NO: 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26, or a substantially identical protein capableof binding and inhibiting IL-1. A preferred IL-1 antagonist is shown inSEQ ID NO:10.

In a fifth aspect, the invention features a method of treating,inhibiting, or ameliorating Familial Cold Autoinflammatory Syndrome(FCAS) the method comprising administering to a subject in need aninterleukin 1 (IL-1) antagonist. In a preferred embodiment, the IL-1antagonist is a fusion protein capable of trapping IL-1. In a specificembodiment, the IL-1 antagonist is the fusion protein shown in SEQ IDNO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, or a substantiallyidentical protein capable of binding and inhibiting IL-1. A preferredIL-1 antagonist is shown in SEQ ID NO:10.

In a sixth aspect, the invention features a method of treating,inhibiting, or ameliorating familial mediterranean fever (FMF), themethod comprising administering to a subject in need an interleukin 1(IL-1) antagonist. In a preferred embodiment, the IL-1 antagonist is afusion protein capable of trapping IL-1. In a specific embodiment, theIL-1 trap is the fusion protein shown in SEQ ID NO:4, 6, 8, 10,12, 14,16, 18, 20, 22, 24, 26, or a substantially identical protein capable ofbinding and inhibiting IL-1. A preferred IL-1 trap is shown in SEQ IDNO:10.

In a seventh aspect, the invention features a method of treating,inhibiting, or ameliorating systemic onset juvenile idiopathic arthritis(Still's Disease), the method comprising administering to a subject inneed an interleukin 1 (IL-1) antagonist. In a preferred embodiment, theIL-1 antagonist is a fusion protein capable of trapping IL-1. In aspecific embodiment, the IL-1 antagonist is the fusion protein shown inSEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, or asubstantially identical protein capable of binding and inhibiting IL-1.A preferred IL-1 trap is shown in SEQ ID NO:10.

In an eighth aspect, the invention features a method of treating,inhibiting, or ameliorating tumour necrosis factor receptor-associatedperiodic fever syndrome (TRAPS), the method comprising administering toa subject in need an IL-1 antagonist. In a preferred embodiment, theIL-1 antagonist is a fusion protein capable of trapping IL-1. In aspecific embodiment, the IL-1 antagonist is the fusion protein shown inSEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, or asubstantially identical protein capable of binding and inhibiting IL-1.A preferred IL-1 trap is shown in SEQ ID NO:10.

In specific embodiments of the therapeutic method of the invention, thesubject is treated with a combination of a first IL-1-binding fusionprotein trap molecule and a second therapeutic agent. The secondtherapeutic agent may be a second IL-1 antagonist, such as, for example,a second IL-1-binding fusion protein trap, anakinra (Kineret®, Amgen), arecombinant, nonglycosylated form of the human IL-1 receptor antagonist(IL1Ra), or an anti-IL-18 drug such as IL-18BP or a derivative, anIL-18-binding fusion protein trap (an “IL-18 trap”), anti-IL-18,anti-IL-18RI, or anti-IL-18Racp antibodies or antibody fragments. Otherco-therapies include low dose colchine for FMF, aspirin or other NSAIDs,steroids such as prednisolone, methotrexate, low dose cyclosporine A,TNF inhibitors such as Enbrel®, or Humira®, other inflammatoryinhibitors such as inhibitors of caspase-1, p38, IKK1/2, CTLA-4Ig,anti-IL-6 or anti-IL6Ra, etc.

In a ninth aspect, the invention features a therapeutic method oftreating an autoinflammatory disease or condition, comprisingadministering a pharmaceutical composition comprising an IL-1-bindingfusion protein trap and a pharmaceutically acceptable carrier. In oneembodiment, the IL-1-binding fusion protein trap is administered in adose range of 1-20 mg/kg on a weekly basis for a treatment period ofbetween 1 week to one year or more. In another embodiment, a totalIL-1-binding fusion protein is administered in the range of 50-2000 mg,which may be provided in a single dose or in sequential doses over aperiod of time such as a period of weeks or months.

Other objects and advantages will become apparent from a review of theensuing detailed description.

DETAILED DESCRIPTION

Before the present methods are described, it is to be understood thatthis invention is not limited to particular methods, and experimentalconditions described, as such methods and conditions may vary. It isalso to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto be limiting, since the scope of the present invention will be limitedonly to the appended claims.

As used in this specification and the appended claims, the singularforms “a”, “an”, and “the” include plural references unless the contextclearly dictates otherwise. Thus for example, a reference to “a method”includes one or more methods, and/or steps of the type described hereinand/or which will become apparent to those persons skilled in the artupon reading this disclosure and so forth.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. All publications mentioned herein areincorporated herein by reference in their entirety.

General Description

Mutations in the gene CIAS1 are now recognized as being responsible forthree rare genetic syndromes: Neonatal Onset Multisystem InflammatoryDisorder (NOMID), Muckle-Wells Syndrome (MWS), and Familial ColdAutoinflammatory Syndrome (FCAS). (Hoffman et al. 2001 Naure 29:301-305;Feldmann et al. 2002 Am J Hum Genet 71:198-203; Aksentijevich et al.2002 Arthritis Rheum 46:3340-3348). In aggregate, these conditions areknown as “CAPS”, an acronym for “CIAS1 Associated Periodic Syndromes”.CAPS disorders are exceedingly rare; with approximately 200-300 adultsand children in the U.S. with FCAS and significantly fewer adults withNOMID or MWS known to have these conditions. The rarity of theseconditions, particularly NOMID and MWS, are probably due to effects ofdisease severity on survival or reproductive fitness.

CAPS are inherited in an autosomal dominant manner, with a sporadic orfamilial pattern. CIAS1 encodes a protein called NALP3 that is acomponent of the “inflammasome”, a subcellular enzyme complex thatregulates the activity of caspase 1. Caspase 1 is the enzyme thatcleaves the inactive pro-form of the proinflammatory cytokine, IL-1,into its biologically active form (Agostini et al. 2004 supra).Mutations in CIAS1 lead to increased production of IL-1 and numerouspathological consequences (Aksentijevich et al. 2002 supra). IL-1strongly induces the production of acute phase reactants in the liver,such as C-reactive protein (CRP) and serum amyloid A (SAA).

The genetics of CAPS are interesting in that there can be a number ofdifferent point mutations in CIAS1 associated with these syndromes(Sarrauste de Menthiere et al. 2003 Nucleic Acids Res 31:282-285;Aksentijevich et al. 2002 supra). Some of these mutations are associatedwith only one syndrome; others two. For example, some mutations may beassociated with FCAS as well as MWS; other mutations may be associatedwith MWS and NOMID. Approximately 50% of patients with NOMID do not havea recognized mutation in the coding region of CIAS1. In these patients,the disease may be due to an as-yet-unrecognized mutation in aregulatory region or protein of CIAS1, or in another gene encoding aclosely-related protein in this pathway. FCAS is more geneticallyhomogeneous than NOMID; almost all patients with FCAS share a commonmutation (Sarrauste de Menthiere et al. 2003 supra; Hoffman et al. 2001supra).

CAPS disorders share common clinical features and present as a spectrumof clinical severity. NOMID is the most seriously disabling, MWSsomewhat less so and FCAS is the least severe. CAPS disorders haveoverlapping features and there are individuals and kindred with uniqueconstellations of signs and symptoms. Features common to all theseconditions include fevers, urticaria-like rash, arthritis or arthralgia,myalgia, malaise, and conjunctivitis. However, the spectrum of symptomsfor any patient with a CAPS disorder may differ from that of anotherpatient with the same disorder. A universal feature of active CAPSdisease is laboratory test elevation of acute phase reactants, such asCRP, SAA, and/or erythrocyte sedimentation rate (ESR).

In NOMID, chronic aseptic meningitis may lead to mental retardation andthese patients may also suffer disfiguring and disabling bony overgrowthat the epiphyses and patellae. These patients may also suffer blindnessdue to optic nerve atrophy that results from increased intracranialpressure. MWS and NOMID are commonly associated with severe inflammationthat may include the auditory system, meninges, and joints. Thesepatients may suffer daily high spiking fevers and a chronic rash thatfrequently changes in distribution and intensity. Patients may sufferhearing loss or deafness. Conjunctivitis and papilledema are frequentlyobserved. Amyloidosis may develop and lead to renal failure due tochronic inflammation and overproduction of acute phase reactants(particularly SM). MWS is also known as “amyloidosis-deafness syndrome”.

The clinical signs and symptoms of FCAS are induced by exposure tomodestly cold air (e.g., seasonal temperature changes, airconditioning). Patients may have frequent (sometimes daily) episodes ofa painful or pruritic rash, fever, fatigue, malaise, headache, nausea,and thirst during cold months or in locations where air conditioning isprevalent. In many locales, this may include most work places. FCAS is asource of frequent pain to patients and may restrict their employment,social, and recreational opportunities. Up to 2% of patients with FCASdevelop amyloidosis, a life-threatening condition. This frequency issubstantially higher than the rate of amyloidosis in the generalcommunity. The genetics and natural history of FCAS are described indetail Hoffman et al. 2001 Nature 29:301-305 and Hoffman et al. 2001 JAllergy din Immunol 108:615-620, which publications are hereinspecifically incorporated by reference in their entirety.

Definitions

By the term “blocker”, “inhibitor”, or “antagonist” is meant a substancethat retards or prevents a chemical or physiological reaction orresponse. Common blockers or inhibitors include but are not limited toantisense molecules, antibodies, antagonists and their derivatives. Morespecifically, an example of an IL-1 blocker or inhibitor is an IL-1antagonist including, but not limited to, an IL-1 fusion protein trapantagonist, which binds and inhibits IL-1.

By the term “therapeutically effective dose” is meant a dose thatproduces the desired effect for which it is administered. The exact dosewill depend on the purpose of the treatment, and will be ascertainableby one skilled in the art using known techniques (see, for example,Lloyd (1999) The Art, Science and Technology of PharmaceuticalCompounding).

By the term “substantially identical” is meant a protein sequence havingat least 95% identity to an amino acid sequence selected from the groupconsisting of the amino acid sequences SEQ ID NOs: 4, 6, 8, 10, 12, 14,16, 18, 20, 22, 24, and 26, and capable of binding IL-1 and inhibitingthe biological activity of IL-1.

The term “identity” or “homology” is construed to mean the percentage ofamino acid residues in the candidate sequence that are identical withthe residue of a corresponding sequence to which it is compared, afteraligning the sequences and introducing gaps, if necessary to achieve themaximum percent identity for the entire sequence, and not consideringany conservative substitutions as part of the sequence identity. NeitherN- or C-terminal extensions nor insertions will be construed as reducingidentity or homology. Methods and computer programs for the alignmentare well known in the art. Sequence identity may be measured usingsequence analysis software (e.g., Sequence Analysis Software Package,Genetics Computer Group, University of Wisconsin Biotechnology Center,1710 University Ave., Madison, Wis. 53705). This software matchessimilar sequences by assigning degrees of homology to varioussubstitutions, deletions, and other modifications.

IL-1-Binding Fusion Protein Trap Antagonists

Interleukin-1 (IL-1) traps are multimers of fusion proteins containingIL-1 receptor components and a multimerizing component capable ofinteracting with the multimerizing component present in another fusionprotein to form a higher order structure, such as a dimer. TheIL-1-binding fusion proteins useful in the methods of the inventioninclude two distinct receptor components that bind a single cytokine,resulting in the generation of antagonists with dramatically increasedaffinity over that offered by single component reagents. TheIL-1-binding fusion protein traps are comprised of the extracellulardomain of human IL-1 R Type I (IL-1RI) or Type II (IL-1RII) followed bythe extracellular domain of human IL-1 Accessory protein (IL-1AcP),followed by a multimerizing component. In a preferred embodiment, themultimerizing component is an immunoglobulin-derived domain, such as,for example, the Fc region of human IgG, including part of the hingeregion, the CH2 and CH3 domains. An immunoglobulin-derived domain may beselected from any of the major classes of immunoglobulins, includingIgA, IgD, IgE, IgG and IgM, and any subclass or isotype, e.g. IgG1,IgG2, IgG3 and IgG4; IgA-1 and IgA-2. Alternatively, the IL-1-bindingfusion proteins useful in the method of the invention are comprised ofthe extracellular domain of human IL-1AcP, followed by the extracellulardomain of human IL-1RI or IL-1RII, followed by a multimerizingcomponent. For a more detailed description of the IL-1-binding fusionprotein traps, see WO 00/18932, which publication is herein specificallyincorporated by reference in its entirety. Preferred IL-1 antagonistshave the amino acid sequence shown in SEQ ID NOs: 2, 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, and 26, or a substantially identical protein atleast 95% identity to a sequence of SEQ ID NO: 4, 6, 8, 10, 12, 14, 16,18, 20, 22, 24, or 26, and capable of binding and inhibiting IL1.

Treatment Population

The therapeutic methods of the invention are useful for treatingindividuals affected with CIAS-1 mutation disorders (NOMID, MWS, FCAS),FMF, TRAPS, or Still's Disease. Commonly accepted diagnostic criteriafor CIAS-1 mutation associated disease (NOMID, MWS, FCAS), FamilialMediterranean Fever, or Still's Disease (adult- or juvenile-onset) areknow to those skilled in the art. In the case of patients diagnosed withFMF, the therapeutic method of the invention may be particularly usefulfor those with disease refractory to therapy with colchicine.

Methods of Administration

The invention provides methods of treatment comprising administering toa subject an effective amount of an agent of the invention. In apreferred aspect, the agent is substantially purified (e.g.,substantially free from substances that limit its effect or produceundesired side-effects).

Various delivery systems are known and can be used to administer anagent of the invention, e.g., encapsulation in liposomes,microparticles, microcapsules, recombinant cells capable of expressingthe compound, receptor-mediated endocytosis (see, e.g., Wu and Wu, 1987,J. Biol. Chem. 262:4429-4432), construction of a nucleic acid as part ofa retroviral or other vector, etc. Methods of introduction can beenteral or parenteral and include but are not limited to intradermal,intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal,epidural, and oral routes. The compounds may be administered by anyconvenient route, for example by infusion or bolus injection, byabsorption through epithelial or mucocutaneous linings (e.g., oralmucosa, rectal and intestinal mucosa, etc.) and may be administeredtogether with other biologically active agents. Administration can besystemic or local. In addition, it may be desirable to introduce thepharmaceutical compositions of the invention into the central nervoussystem by any suitable route, including intraventricular and intrathecalinjection; intraventricular injection may be facilitated by anintraventricular catheter, for example, attached to a reservoir, such asan Ommaya reservoir. Pulmonary administration can also be employed,e.g., by use of an inhaler or nebulizer, and formulation with anaerosolizing agent.

In a specific embodiment, it may be desirable to administer thepharmaceutical compositions of the invention locally to the area in needof treatment; this may be achieved, for example, and not by way oflimitation, by local infusion during surgery, topical application, e.g.,by injection, by means of a catheter, or by means of an implant, saidimplant being of a porous, non-porous, or gelatinous material, includingmembranes, such as sialastic membranes, fibers, commercial skinsubstitutes or angioplasty balloons or stents.

In another embodiment, the active agent can be delivered in a vesicle,in particular a liposome (see Langer (1990) Science 249:1527-1533). Inyet another embodiment, the active agent can be delivered in acontrolled release system. In one embodiment, a pump may be used (seeLanger (1990) supra). In another embodiment, polymeric materials can beused (see Howard et al. (1989) J. Neurosurg. 71:105). In anotherembodiment where the active agent of the invention is a nucleic acidencoding a protein, the nucleic acid can be administered in vivo topromote expression of its encoded protein, by constructing it as part ofan appropriate nucleic acid expression vector and administering it sothat it becomes intracellular, e.g., by use of a retroviral vector (see,for example, U.S. Pat. No. 4,980,286), or by direct injection, or by useof microparticle bombardment (e.g., a gene gun; Biolistic, Dupont), orcoating with lipids or cell-surface receptors or transfecting agents, orby administering it in linkage to a homeobox-like peptide which is knownto enter the nucleus (see e.g., Joliot et al., 1991, Proc. Natl. Acad.Sci. USA 88:1864-1868), etc. Alternatively, a nucleic acid can beintroduced intracellularly and incorporated within host cell DNA forexpression, by homologous recombination.

Combination Therapies

In numerous embodiments, the IL-1 antagonists useful in the methods ofthe present invention may be administered in combination with one ormore additional compounds or therapies. Combination therapy may besimultaneous or sequential. The IL-1-binding fusion proteins of theinvention may be combined with, for example, TNF-inhibiting agents suchas etanercept (Enbrel®, Amgen), infliximab (Remicade®, Centocor),Humira® (Abbott), thalidomide, steroids, anakinra (Kinaret®, Amgen), orcolchicine. Colchicine is a mainstay of therapy for subjects with FMF;in this study, subjects will not be removed from treatment with thismedication. For Still's Disease (and classical autoinflammatorydiseases), compounds such as methotrexate, cyclosporine, chlorambucil,cyclophosphamide (DMARDs) have been used as monotherapy or incombination with no consistent response. Some subjects respond to highdoses of steroids. DMARDs, and more recently anti-TNF agents have beenused with variable success. The IL-1-binding fusion proteins of theinvention may also be combined with anti-IL-18 drugs, such as forexample, IL-18BP or a derivative, an IL-18-binding fusion protein,anti-IL-18, anti-IL-18RI, or anti-IL-18Racp. Other co-therapies includelow dose colchine for FMF, aspirin or other NSAIDs, steroids such asprednisolone, methotrexate, low dose cyclosporine A, TNF inhibitors suchas Enbrel®, or Humira®, other inflammatory inhibitors such as inhibitorsof caspase-1, p38, IKK1/2, CTLA4Ig, anti-IL-6 or anti-IL6Ra, etc.

Pharmaceutical Compositions

The present invention also provides pharmaceutical compositions. Suchcompositions comprise a therapeutically effective amount of an activeagent, and a pharmaceutically acceptable carrier. The term“pharmaceutically acceptable” means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopeia orother generally recognized pharmacopeia for use in animals, and moreparticularly, in humans. The term “carrier” refers to a diluent,adjuvant, excipient, or vehicle with which the therapeutic isadministered. Such pharmaceutical carriers can be sterile liquids, suchas water and oils, including those of petroleum, animal, vegetable orsynthetic origin, such as peanut oil, soybean oil, mineral oil, sesameoil and the like. Suitable pharmaceutical excipients include starch,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, sodium stearate, glycerol monostearate, talc, sodium chloride,dried skim milk, glycerol, propylene, glycol, water, ethanol and thelike. The composition, if desired, can also contain minor amounts ofwetting or emulsifying agents, or pH buffering agents. Thesecompositions can take the form of solutions, suspensions, emulsion,tablets, pills, capsules, powders, sustained-release formulations andthe like. The composition can be formulated as a suppository, withtraditional binders and carriers such as triglycerides. Oral formulationcan include standard carriers such as pharmaceutical grades of mannitol,lactose, starch, magnesium stearate, sodium saccharine, cellulose,magnesium carbonate, etc. Examples of suitable pharmaceutical carriersare described in “Remington's Pharmaceutical Sciences” by E. W. Martin.

In a preferred embodiment, the composition is formulated in accordancewith routine procedures as a pharmaceutical composition adapted forintravenous administration to human beings. Where necessary, thecomposition may also include a solubilizing agent and a local anestheticsuch as lidocaine to ease pain at the site of the injection. Where thecomposition is to be administered by infusion, it can be dispensed withan infusion bottle containing sterile pharmaceutical grade water orsaline. Where the composition is administered by injection, an ampouleof sterile water for injection or saline can be provided so that theingredients may be mixed prior to administration.

The active agents of the invention can be formulated as neutral or saltforms. Pharmaceutically acceptable salts include those formed with freeamino groups such as those derived from hydrochloric, phosphoric,acetic, oxalic, tartaric acids, etc., and those formed with freecarboxyl groups such as those derived from sodium, potassium, ammonium,calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.

The amount of the active agent of the invention which will be effectivein the treatment of delayed-type hypersensitivity can be determined bystandard clinical techniques based on the present description. Inaddition, in vitro assays may optionally be employed to help identifyoptimal dosage ranges. The precise dose to be employed in theformulation will also depend on the route of administration, and theseriousness of the condition, and should be decided according to thejudgment of the practitioner and each subject's circumstances. However,suitable dosage ranges for intravenous administration are generally upto about 2 grams of active compound. Effective doses may be extrapolatedfrom dose-response curves derived from in vitro or animal model testsystems.

For systemic administration, a therapeutically effective dose can beestimated initially from in vitro assays. For example, a dose can beformulated in animal models to achieve a circulating concentration rangethat includes the IC₅₀ as determined in cell culture. Such informationcan be used to more accurately determine useful doses in humans. Initialdosages can also be estimated from in vivo data, e.g., animal models,using techniques that are well known in the art. One having ordinaryskill in the art could readily optimize administration to humans basedon animal data.

Dosage amount and interval may be adjusted individually to provideplasma levels of the compounds that are sufficient to maintaintherapeutic effect. In cases of local administration or selectiveuptake, the effective local concentration of the compounds may not berelated to plasma concentration. One having skill in the art will beable to optimize therapeutically effective local dosages without undueexperimentation.

The amount of compound administered will, of course, be dependent on thesubject being treated, on the subject's weight, the severity of theaffliction, the manner of administration, the frequency ofadministration and the judgment of the prescribing physician. Thetherapy may be repeated intermittently while symptoms are detectable oreven when they are not detectable. The therapy may be provided alone orin combination with other drugs.

Kits

The invention also provides an article of manufacturing comprisingpackaging material and a pharmaceutical agent contained within thepackaging material, wherein the pharmaceutical agent comprises at leastone IL-1-specific fusion protein of the invention and wherein thepackaging material comprises a label or package insert which indicatesthat the IL-1-specific fusion protein can be used for treating anautoinflammatory disease or condition.

Other features of the invention will become apparent in the course ofthe following descriptions of exemplary embodiments which are given forillustration of the invention and are not intended to be limitingthereof.

EXAMPLES

The following example is put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the methods and compositions of the invention, and are notintended to limit the scope of what the inventors regard as theirinvention. Efforts have been made to ensure accuracy with respect tonumbers used (e.g., amounts, temperature, etc.) but some experimentalerrors and deviations should be accounted for. Unless indicatedotherwise, parts are parts by weight, molecular weight is averagemolecular weight, temperature is in degrees Centigrade, and pressure isat or near atmospheric.

Example 1 Effect of IL-1 Trap on Human Autoinflammatory Disease

An initial study is conducted with 15 adult subjects suffering fromdiseases known to respond to IL-1 blockade (NOMID/MWS/FCAS) as well assubjects with Adult Still's disease and colchicine-resistant FMF.Subjects are screened for eligibility, clinical symptoms determined,active disease confirmed and baseline blood is drawn on approximately 3occasions one week apart to determine baseline levels of inflammation. Acareful, complete standardized history and physical exam is performed,appropriate for the disease under study to assure uniform datacollection on every subject. Vital signs and weight is obtained at eachvisit. The clinical data is based on a detailed questionnaire includingall the reported clinical manifestations. The following evaluationprocedures pertain specifically to CIAS-1 mutation associated disordersand are performed as clinically indicated: dermatological evaluation;ophthalmologic evaluation; ear/nose/throat evaluation; neurologyevaluation; lumbar puncture; head MRI; radiographs, joint MRI; andpharmacokinetic profiling.

All study subjects receive IL-1-binding fusion protein (SEQ ID NO:10)with a dosing regimen of 100 mg once a day for 3 consecutive days, aregimen expected to provide 2-4 weeks of significant IL-1 inhibitoryactivity. The primary outcomes are measured during this period andinclude drug safety, clinical efficacy analysis, and the change inselected biomarkers of inflammation (e.g., acute phase reactants such asCRP, serum amyloid A, and ESR) at Day 10 following initiation oftreatment with IL-1 trap. If a favorable response is observed at Day 10,subjects are monitored at predefined timepoints (with no furthertreatment) until return of signs and symptoms (flare). Upon flare,subjects are eligible for entry into an extension phase that entailsre-treatment with the loading regimen (100 mg/day IL-1 trap for threeconsecutive days) followed by once-weekly dosing with 100 mg IL-1 trapfor up to one year.

Based on the Investigator's clinical judgment, an IL-1 trap doseescalation regimen may be implemented if, after 4 weeks of dosing in theextension phase at 100 mg/week, a subject's Month 1 acute phase reactantlevels have not normalized (CRP>0.5 mg/dL and/or SAA>10 mg/L) orescalation is warranted based on persistent signs and/or symptoms ofdisease. The first dose escalation level may be 160 s.c. once weekly.Subjects will be observed for 4 weeks; if criteria for dose escalationare still met, then the dose may be raised to 320 mg s.c. once-weekly.

Preliminary Results. Four subjects with CAPS were initially enrolled.Results indicated that all subjects experienced rapid and extensiveimprovement in inflammatory signs and symptoms upon treatment with IL-1trap (SEQ ID NO:10), including improvement in both patient- andphysician-reported disease manifestation. Major declines in inflammatorybiomarkers, such as CRP and SAA were also observed. Signs and symptomsreturned within a median of 21 days (range 9-26) of initial dosing andthen responded promptly to re-treatment. Table 1 provides a summary ofthe daily diary scores, acute phase reactants and clinical assessments(t Performed on 3 patients; * statistically significant difference fromprevious time point at p<0.1 level; ** statistically significantdifference from previous time point at p<0.05 level). The Physician andPatient global assessment VAS scores mirrored the changes in the acutephase reactants (SM, CRP and ESR) at baseline, at the time of flare, andat a time point designated as reflecting maximal efficacy. TABLE 1Baseline Maximal Efficacy Flare median (range) median (range) median(range) Daily Diary Score  6.06 (2.2-7.56)  1.67 (0-3.3)*  4.5 (2-7.33)Acute phase reactants SAA (mg/L)    96 (16.1-468)  8.25 (2-19)   84(50-236)‡ CRP (mg/dL)  7.28 (2.32-8.65)  0.72 (0.07-1.15)**  2.93(0.076-6.21) ESR (mm/hr)  56.67 (22-92)    24 (7-45)**   34 (11-70)*Blood Count WBC  15.28 (9.33-19.4)  7.58 (7.21-9.9)**  8.48 (6.34-11.47)Hgb  12.95 (8.1-14.7)  13.3 (8.2-15.6)*  13.1 (7.9-14.57) Plt  356.5(291-445.5) 303.25 (240-377)**   291 (257-359.3) Questionnaires‡Physician global VAS  6.85 (4.1-6.95)   0.2 (0.2-2.6)  3.3 (3.1-3.5)(cm) Patient global VAS (cm)   5.2 (3.95-6.9)   1.1 (0.95-3.05)**  3.6(3.1-6.45)** Fatigue VAS (cm)  5.55 (3.25-8)  1.15 (0.5-3.9)  6.6(3.15-6.9) Pain VAS (cm)  7.55 (3.6-7.7)  0.95 (0.2-1.05)*  4.1(0.5-6.55) SF-36 Physical Health  44.38 (42.5-47.5)  50.63 (33.75-92.5)41.56 (35-69.4) SF-36 Mental Health 41.625 (28.5-57.8)  75.88 (55-96) 39.6 (37-57)

1. A method of treating, inhibiting, or ameliorating an autoinflammatorydisorder, disease, or condition in a subject in need thereof, comprisingadministering to a subject a therapeutic amount of an interleukin 1(IL-1) fusion protein antagonist, wherein the autoinflammatory disorder,disease, or condition is treated, inhibited, or ameliorated, wherein theIL-1 fusion protein antagonist comprises two IL-1 receptor componentsand a multimerizing component.
 2. The method of claim 1, wherein thefusion protein antagonist comprises an amino acid sequence selected fromthe group of amino acid sequences shown in SEQ ID NO: 4, 6, 8, 10, 12,14, 16, 18, 20, 22, 24, 26, or a substantially identical having at least95% identity to the sequence shown in SEQ ID NO: 4, 6, 8, 10, 12, 14,16, 18, 20, 22, 24, 26 and capable of binding and inhibiting IL-1. 3.The method of claim 2, wherein the fusion protein antagonist comprisesSEQ ID NO:10.
 4. The method of claim 1, wherein the subject is a humanadult or child diagnosed with Neonatal Onset Multisystem InflammatoryDisorder (NOMID/CINCA), Muckle-Wells Syndrome (MWS), Familial ColdAutoinflammatory Syndrome (FCAS), familial mediterranean fever (FMF),tumor necrosis factor receptor-associated periodic fever syndrome(TRAPS), or systemic onset juvenile idiopathic arthritis (Still'sDisease).
 5. The method of claim 1, wherein administration issubcutaneous, intramuscular, intravenous, topical, transdermal or oral.6. The method of claim 1, wherein a therapeutically effective amount isbetween 1-20 mg/kg.
 7. The method of claim 1, further comprisingadministering a second therapeutic agent or agents, wherein the secondtherapeutic agent is one or agents selected from the group consisting ofan IL-1 fusion protein which is different from the first agent,etanercept (Enbrel®, Amgen), infliximab (Remicade®, Centocor), Humira®(Abbott), thalidomide, a steroid, anakinra (Kinaret®, Amgen),colchicine, IL-18BP or a derivative, an IL-18-binding fusion protein,anti-IL-18, anti-IL-18RI, anti-IL-18Racp, aspirin, prednisolone,methotrexate, cyclosporine A, caspase-1, p38, IKK1/2, CTLA-4Ig,anti-IL-6 and anti-IL6Ra.
 8. A method of treating, inhibiting, orameliorating an autoinflammatory disorder associated with mutations inCIAS-1 in a subject in need thereof, comprising administering to asubject in need a therapeutic amount of an interleukin 1 (IL-1)antagonist, wherein the autoinflammatory disorder, disease, or conditionis treated, inhibited, or ameliorated.
 9. The method of claim 8, whereinthe autoinflammatory disorder associated with mutations in CIAS-1 is oneof Neonatal Onset Multisystem Inflammatory Disorder (NOMID/CINCA),Muckle-Wells Syndrome (MWS), Familial Cold Autoinflammatory Syndrome(FCAS).
 10. The method of claim 8, wherein the IL-1 antagonist is afusion protein having an amino acid sequence selected from the group ofamino acid sequences shown in SEQ ID NO: 4, 6, 8, 10, 12, 14, 16, 18,20, 22, 24, 26, or a substantially identical having at least 95%identity to the sequence shown in SEQ ID NO: 4, 6, 8, 10,12, 14, 16, 18,20, 22, 24, 26 and capable of binding and inhibiting IL-1.
 11. Themethod of claim 10, wherein the fusion protein comprises SEQ ID NO:10.12. A method of treating, inhibiting, or ameliorating anautoinflammatory disorder associated with mutations in CIAS-1 in asubject in need thereof, comprising administering to a subject in need atherapeutic amount of an interleukin 1 (IL-1) antagonist, wherein theautoinflammatory disorder, disease, or condition is treated, inhibited,or ameliorated, wherein the IL-1 antagonist is a fusion proteincomprising two IL-1 receptor components and a multimerizing component.13. The method of claim 12, wherein the fusion protein comprises SEQ IDNO:10.
 14. The method of claim 12, further comprising administering asecond therapeutic agent.
 15. The method of claim 14, wherein the secondtherapeutic agent is selected from the group consisting of an IL-1inhibiting fusion protein, etanercept (Enbrel®, Amgen), infliximab(Remicade®, Centocor), Humira® (Abbott), thalidomide, a steroid,anakinra (Kinaret®, Amgen), colchicine, IL-18BP or a derivative, anIL-18 receptor fusion protein, anti-IL-18, anti-IL-18RI, anti-IL-18Racp,aspirin prednisolone, methotrexate, cyclosporine A, caspase-1, p38,IKK1/2, CTLA-4Ig, anti-IL-6 and anti-IL6Ra.
 16. A method of treating,inhibiting, or ameliorating an autoinflammatory disorder associated withmutations in CIAS-1 in a subject in need thereof, comprisingadministering to a subject in need a therapeutic amount of aninterleukin 1 (IL-1) antagonist comprising the amino acid sequence ofSEQ ID NO:10, wherein the autoinflammatory disorder, disease, orcondition is treated, inhibited, or ameliorated.